OBJECTIVE: To study DNA quantification and STR typing of samples pre-treated with pyramidon. METHODS: The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accordance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology. RESULTS: In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples. CONCLUSION Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extraction is the best method for STR profiling and DNA extraction.
Aminopyrine/pharmacology* ; Blood Stains ; DNA/isolation & purification* ; DNA Fingerprinting ; Forensic Medicine ; Humans ; Polymerase Chain Reaction ; Reproducibility of Results ; Specimen Handling

OBJECTIVETo study value of velocity vector imaging (VVI) in assessment of normal left ventricular diastolic function and the corresponding reference values.

METHODSNinety-seven healthy subjects were selected by combined clinical, ultrasound, and NT-proBNP examinations. Using a Siemens Acuson Sequoia C512 echocardiograph, VVI was adopted to examine the myocardial early diastolic velocity (E) of the septal, lateral, anterior, inferior, anterior septum, and posterior wall of the left ventricle at the level of mitral annulus. The images were analyzed for VVI and the mean diastolic velocity (E(m)) and hence the E/E(m) ratio was calculated based on the offline workstation interface.

RESULTSThe reference range of E/E(m) ratio derived from the data of the 97 healthy subjects was (unilateral boundaries with 95% limit) and (1, 22.935), was (1, 22.300) in male subjects and (1, 24.766) in female subjects. The reference E/E(m) range was (0, 22.413) in male subjects under 50 years of age, (1, 24.766) in female subjects under 50 years, (1, 22.300) in male subjects over 50 years, and (1, 24.766) in female subjects over 50 years.

CONCLUSIONVVI is a good method for non-invasive evaluation of the left ventricular diastolic function and provides an accurate and reliable means for clinical assessment of the left ventricular diastolic function.

Adolescent ; Adult ; Aged ; Blood Flow Velocity ; physiology ; Diastole ; physiology ; Echocardiography ; methods ; Female ; Humans ; Male ; Middle Aged ; Reference Values ; Ventricular Function, Left ; physiology ; Young Adult

OBJECTIVEAllogeneic marrow transplantation is a curative therapy for thalassemia, but no more than 30% of patients have HLA-indentical sibling marrow donor. The selection of alternative donors of unrelative marrow and the study on the probability of treating thalassemia major with unrelated donor bone marrow transplantation are of importance.

METHODSNine children with thalassemia were included in the study, and their gene mutational type were homozygote of thalassemia and double heterozygote, respectively. All of them were finally diagnosed of thalassemia major, and treated with unrelated donor bone marrow transplantation. To high-resolution HLA typing, two patients were matched, five had one unmatched isoform and two had two unmatched isoforms. The erythrocyte blood type was not matched in six patients. The preparative regimen included busulfan (oral use, 16 mg/kg, divided for 4 days), cyclophosphamide (intravenous use, 200 mg/kg, divided for 4 days), antithymocyte immunoglobulin (intravenous use, 30 mg/kg, divided for 3 days), and fludarabine (intravenous use, 125 mg/m(2), divided for 3 days). Ciclosporin A and methotrexate were used for graft-versus-host disease (GVHD) prophylaxis.

RESULTSAll patients had allergen reactions. One had hypotension. Five patients experienced I degrees approximately III degrees acute GVHD in the skin, while one had II degrees acute GVHD in liver. One patient had III degrees GVHD of intestines and gradually developed chronic GVHD in the skin, lungs and brain. One patient died of pulmonary hemorrhage. The duration when peripheral blood neutrophil count exceeded 0.5 x 10(9)/L was 12 - 26 days. The recovery time of WBC was as long as 23 - 110 days. Thrombocytes exceeded 50 x 10(9) within 61 approximately 142 days. The time when hemoglobin reached 100 g/L varied from 23 to 116 days. The last blood transfusion was on 13 - 62 days. Eight patients were fully grafted, while one was not grafted. During the 6 - 24 months of follow-up, seven patients' genotype of thalassemia major became normal. The erythrocyte blood type of five patients also changed into the same as that of donor. The hemoglobin was kept over 110 g/L without blood transfusion.

CONCLUSIONThe transplantation of unrelated donor bone marrow for thalassemia major was successful. Unrelated donor bone marrow transplantation could cure thalassemia major, which expanded the marrow donor source for the transplantation of thalassemia major.

ABO Blood-Group System ; Bone Marrow Transplantation ; adverse effects ; Child ; Child, Preschool ; Disease-Free Survival ; Female ; Follow-Up Studies ; Graft Rejection ; Graft Survival ; Histocompatibility Testing ; Humans ; Infant ; Male ; Transplantation Tolerance ; Transplantation, Homologous ; adverse effects ; Treatment Outcome ; beta-Thalassemia ; diagnosis ; therapy

Background Since the measurement method establishment of serum ferritin abroad in early period of theseventies, the iron deficiency had been divided into two types: the non-anemia and anemia types. In orderto go step further studies, we must ertablish the bemoglobin targets of the two types. Methods One hurdred and fifty-two children in experimontal group, from 6 to 7 years old, and allcome from Qinghai province. There are 29 children in Xining city, 24 in Guide, 26 in Gongbe, 40 in Gui-nan and 33 in Maduo countics. There are 36 health children aged from 6 to 7 years old in the controlgroup, and all comes from Beijing. The Hb, RBC, HCT, HCTW and FEP wcre determined. Results The three targets correlating with Hb (Hb, MCH and MCHC); correlating with RBC (RBC,HCT and MCV); the two targets correlating with RBC_weight (HCTW and CMCW) and correlating withFEP of RBC(FEP and MCEP) have very significant difference between experimental group and control group. Conclusion The determination values of the 10 targets are not same in children in different districts,and the values of all the target: are increased on different degree along with the increase in altitude of ele-vation. There is very important significance on the studies of iron deficiency and altitude hypoxia to establish the normal values of the 10 targets.

Objective To determine the current status and influence hctors of reporting quality of the Chinese meta-analysis on stomatology.Methods A comprehensive electronic search was carried out through Chinese BioMedical Literature Database(CBM),VIP Databage for Chinese Technical Periodicals (VIP)and China National Knowledge Infrastructure(CNKI)and a hand searching was also performed through 19 stomatologieal journals in Chinese to identify meta-analysis on stomatology.Two reviewers took responsibility for study inclusion,data extraction and reporting quality assessment with preferred reporting items for systematic reviews and meta-analysis(PRISMA)in duplicate and any disagreement was resolved by discussion.Results A total of 34 meta-analysis on stomatology were eligible,most of them were ou oral medicine and oral and maxillofacial surgery,and mainly focusing on etiology,prevention and treatment of oral diseases.The number of the meta-analysis increased during recent years.Reporting quality of the meta-analysis was not hish and the PRISMA scored(13.6±4.2).The main factors that influenced the reporting quality of meta-analysis were published on evidence-based medicine journals(adjusted β=0.53,t=4.15,showed that this outcome was stable.Conclusions Reposing quality of the Chinese meta-analysis on stomatology is low.To provide sufficient evidence to the clinicians and promote the development of evidencebased dentistry in China,experts on stomatology should study the knowledge of evidence-based medicine and comply with PRISMA statement when producing the meta-analysis.

This study was aimed to investigate the effect of CD44 gene silence on the drug resistance and biologic activity of human multidrug resistant leukemia cell line K562/A02. The oligonucleotides of CD44 gene were designed according to related data of GenBank, double-stranded DNA was produced by annealing, and was inserted into pGCsilencerU6/Neo/GFP vector. The resultant recombinant plasmid pGCsiRNA-CD44 was transfected into K562/A02 cell line. Expressions of CD44, mdr-1 and blc-2 mRNA were assayed by real time RT-PCR. The 50% inhibitory concentration (IC50) of doxorubicin (ADM) for K562/A02 cell line was determined by MTT method. Cell cycle was determined by flow cytometry. The morphology of apoptotic cells was examined by Hochst 33258 staining. The results indicated that the siRNA eukaryotic plasmid directing at CD44 gene could effectively silence the CD44 gene of K562/A02 cells; as compared with control group, the CD44 expression in K562/A02 cells transfected with 4 pGCsiRNA-CD44 plasmids was obviously inhibited, while the inhibition of CD44 expression in cells transfected with siCD44-1 was strongest. After being transfected with pGCsiRNA-CD44, the expression of CD44 mRNA in K562/A02 cells reduced by 64.1% (p<0.05), at the same time the expression of mdr-1 and bcl-2 mRNA in pGCsiRNA-CD44-transfected K562/A02 cells reduced by 25.6% and 50.8% respectively. IC50 of K562/A02 cells after transfection decreased to (8.77+/-1.63) microg/ml and was obviously lower than that of control (17.97+/-1.61) microg/ml (p<0.01). After transfection for 48 hours, the ratio of K562/A02 cells in G0/G1 increased by 10.7%, and the cells displayed karyopyknosis, nuclear margination and apoptotic bodies. It is concluded that the siRNA plasmid specifically targeting CD44 gene can remarkably down-regulate the expression of CD44 gene, inhibit K562/A02 cell proliferation, induce its apoptosis and effectively reverse the multidrug resistance of K562/A02 cells.
Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Gene Silencing ; Genetic Vectors ; Humans ; Hyaluronan Receptors ; genetics ; K562 Cells ; RNA, Small Interfering

OBJECTIVETo investigate the effect of DNA methylation in combination with histone deacetylase inhibitor on transcription regulation of Ras associated domain family gene 1(RASSF1A) tumor suppressor gene and the molecular biological behaviors in U266 cells.

METHODSThe U266 cells were treated with different doses of 5-Aza-2'-deoxycytidine (5-Aza-CdR) and Valproate (VPA) each alone or in combination. Methylation-specific PCR (MSP) was used to detect CpG island methylation in RASSF1A promoter. Quantitative real-time reverse transcription polymerase chain reaction (RQ-PCR) was used to examine the expression of RASSF1A gene in U266 cells. MTT was used for cell proliferation. Cell apoptosis and cell cycle were analyzed by flow cytometry.

RESULTSThe methylation of RASSF1A gene promoter was detected in U266 cells, while there was little RASSF1A gene expressing in the control group. The demethylation effect could be detected in the 5-Aza-CdR treated and combined treatment groups but no in the VPA group. The expression level of RASSF1A was induced by 5-Aza-CdR in a concentration-dependent manner while VPA had no such effect. The expression level of RASSF1A mRNA was increased significantly in the combined treatment group. Higher growth inhibition and apoptosis effects were found in 5-Aza-CdR and VPA combination group than that in 5-Aza-CdR or VPA alone group (P < 0.05). After treatment with 5-Aza-CdR or VPA alone for 72 h, more cells were arrested in G(0)/G(1) phase as conpared with control group (P < 0.05), and even more cells were so arrested in combined treatment group (P < 0.05).

CONCLUSIONDNA methylation and histone deacetylase inhibitor can synergistically induce demethylation of the RASSF1A gene, re-express RASSF1A gene silenced in U266 cells, inhibit the proliferation of U266 cells and induce cell apoptosis.

Cell Line, Tumor ; CpG Islands ; DNA Methylation ; Gene Expression ; Histone Deacetylase Inhibitors ; Humans ; Promoter Regions, Genetic

Objective To explore the relationship between post-traumatic stress disorder (PTSD) and perceived social support (PSS)ability among middle school students in earthquake-stricken areas after the Wenchuan earthquake accident. Methods A total of 1966 students from three secondary schools of Wenchuan earthquake-stricken areas were evaluated by PTSD Checklist-Civilian Version (PCL-C) and Perceived Social Support Scale (PSSS). Results The overall incidence rate of PTSD was 78.3%, with severe PTSD as 24.38%. Significant differences on the incidence rates of PTSD were found among the students who were in different PSS levels (P<0.05) and significant negative correlation existed between the levels of severity on PTSD and PSS (γ=-0.226, P<0.05). Significant differences on PTSD incidence rates were found among those students who were from different families or out-family PSS levels (P=0.009, P<0.05). Significant negative correlation existed between the severity of PTSD and family or out-family PSS level (γ=-0.176, P<0.05, γ=-0.214, P<0.05). Conclusion Relationships between the incidence rate, severity of PTSD and PSS levels existed among the middle school students in Wenchuan earthquake-stricken areas, with higher PSS, lower incidence rate and lighter severity of PTSD. Psychological intervention for earthquake-stricken students should be carried out.

Waldenström's macroglobulinemia (WM) is one of malignant hematological disease on account of abnormal proliferation of B lymphocyte clone and the pathologic cells of WM possess ability to secrete monoclonal immunoglobulin M. In this study, the diagnosis and morphological characteristics of 2 patients with WM were analyzed. The results showed that a special kind of "foam cells" were found by cytochemical staining examinations in both cases, which displayed characteristics of lymphocytes, but neither monocyte-macrophage nor fatty cells. The periodic acid-Shiff's reaction (PAS) demonstrated strong positive, especially on the inclusion bodies in pathologic cell plasma while the acid phosphatase, and alpha-butanoic acetate esterase stainings, resulted both in negative. In conclusion, the cells found in the two cases reported may be described as gemmy ring-like lymphocyte in morphology, a special subtype of ring-like lymphocyte.
Adult ; Humans ; Male ; Middle Aged ; Waldenstrom Macroglobulinemia ; pathology

OBJECTIVETo assess the value of velocity vector imaging (VVI) and quantitative tissue velocity imaging (QTVI) in assessing left ventricular diastolic function of the dogs with acute myocardial ischemia.

METHODSSix healthy mongrel dogs were subjected to ligation of the left circumflex artery or left anterior descending artery to induce coronary artery stenosis of varying degrees. The mean peak diastolic velocity (Em) of the ventricular walls around the mitral annulus was recorded with VVI or QTVI in the coronary blood flow. The left ventricular end diastolic pressure (LVEDP) was measured with pigtail catheter in the left ventricle.

RESULTSAs the coronary blood flow decreased, LVEDP was gradually increased, and Em measured by VVI or QTVI were also gradually decreased. A good linear correlation was shown between Em measured by VVI or QTVI and LVEDP (r=-0.834, P<0.001, and r=-0.68, P<0.001, respectively). A significant difference was observed in the correlation coefficient between VVI and QTVI (Z=2.625, P=0.0087).

CONCLUSIONVVI and QTVI both provide good noninvasive means for measuring left ventricular diastolic function. VVI, a new echocardiographic modality without angular dependence, is better than QTVI in evaluating left ventricular diastolic function.

Animals ; Disease Models, Animal ; Dogs ; Echocardiography ; methods ; Male ; Myocardial Ischemia ; diagnostic imaging ; physiopathology ; Ventricular Function, Left